Using a system developed in this laboratory that simultaneously measures membrane potential, DeltapH proton motive force, respiration, proton extrusion, and H-+-/O ratios we have studied the interrelation of these parameters of energy transduction during pulses of respiration with succinate as substrate in intact E. coli under a variety of conditions. These studies show how these quantities adjust themselves to changes in external pH, pK+, [0-2], concentrations of ionophores and probes, and the age of the cells. The H-+-/O ratios measured by the initial kinetics of proton extrusion and oxygen uptake rate are significantly higher than those measured by the more traditional integrated or bulk measurements of quantities of protons extruded and oxygen consumed. These higher ratios are not compatible with the Mitchell hypothesis of proton pumping.